Isolation and characterization of two cyclin cDNAs from Pisum sativum L.

In order to investigate the role of cell division in plant development, we isolated two plant genes which encode homologues of animal and yeast cell cycle regulators known as cyclins. Through the use of degenerate primers and the polymerase chain reaction (PCR) we isolated a Pisum sativum sequence which showed homology to the ‘cyclin box°s functional domain found within cyclin proteins. Using this sequence as probe we isolated two different cyclin cDNAs, Pissa;CycA3;1 and Pissa;CycB1;3 from a Pisum sativum L. root tip cDNA library. The deduced amino acid sequences of both cDNAs showed the highest sequence similarity with mitotic cyclins. Analyses of Pissa;CycA3;1 and Pissa;CycB1;3 expression in different tissues, by reverse transcription-polymerase chain reaction (RT-PCR) using primers corresponding to unique regions of their cDNAs, showed their differential expression in relation to cell cycle activity. Furthermore, RT-PCR was used to analyze synchronized root tip cells; results revealed that Pissa;CycA3;1 is preferentially expressed in mid-S (SM) and during late S-G2 (Sl- G2) transition, whereas Pissa;CycB1;3 mRNA is only detectable in Sl and G2 phases